PMID- 29614494 OWN - NLM STAT- MEDLINE DCOM- 20180705 LR - 20190212 IS - 1421-9778 (Electronic) IS - 1015-8987 (Linking) VI - 46 IP - 2 DP - 2018 TI - DEPTOR Deficiency-Mediated mTORc1 Hyperactivation in Vascular Endothelial Cells Promotes Angiogenesis. PG - 520-531 LID - 10.1159/000488619 [doi] AB - BACKGROUND/AIMS: The mechanistic target of rapamycin (mTOR) signaling pathway is essential for angiogenesis and embryonic development. DEP domain-containing mTOR-interacting protein (DEPTOR) is an mTOR binding protein that functions to inhibit the mTOR pathway In vitro experiments suggest that DEPTOR is crucial for vascular endothelial cell (EC) activation and angiogenic responses. However, knowledge of the effects of DEPTOR on angiogenesis in vivo is limited. This study aimed to determine the role of DEPTOR in tissue angiogenesis and to elucidate the molecular mechanisms. METHODS: Cre/loxP conditional gene knockout strategy was used to delete the Deptor gene in mouse vascular ECs. The expression or distribution of cluster of differentiation 31 (CD31), vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1 alpha (HIF-1alpha) were detected by immunohistochemical staining or western blot. Tube formation assay was used to measure angiogenesis in vitro. RESULTS: Deptor knockdown led to increased expression of CD31, VEGF and HIF-1alpha in heart, liver, kidney and aorta. After treatment with rapamycin, their expression was significantly down regulated. In vitro, human umbilical vein endothelial cells (HUVECs) were transfected with DEPTOR-specific small interfering RNA (siRNA), which resulted in a significant increase in endothelial tube formation and migration rates. In contrast, DEPTOR overexpression markedly reduced the expression of CD31, VEGF and HIF-1alpha. CONCLUSIONS: Our findings demonstrated that deletion of the Deptor gene in vascular ECs resulted in upregulated expression of CD31 and HIF-1alpha, and further stimulated the expression of VEGF which promoted angiogenesis, indicating that disruption of normal angiogenic pathways may occur through hyperactivation of the mTORC1/HIF-1alpha/VEGF signaling pathway. CI - (c) 2018 The Author(s). Published by S. Karger AG, Basel. FAU - Ding, Yan AU - Ding Y AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. AD - Department of Endocrinology, Wuhan General Hospital of Guangzhou Command, Wuhan, Hubei Province, Southern Medical University, Guangzhou, China. FAU - Shan, Lanlan AU - Shan L AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Nai, Wenqing AU - Nai W AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Lin, Xiaojun AU - Lin X AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. FAU - Zhou, Ling AU - Zhou L AD - Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Dong, Xiaoying AU - Dong X AD - Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Wu, Hongyuan AU - Wu H AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Xiao, Min AU - Xiao M AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. FAU - Zhou, Xuejuan AU - Zhou X AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. FAU - Wang, Linlin AU - Wang L AD - Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Li, Ting AU - Li T AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. FAU - Fu, You AU - Fu Y AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Lin, Yijun AU - Lin Y AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Jia, Chunhong AU - Jia C AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. FAU - Dai, Meng AU - Dai M AD - From the Department of Health Management, Nanfang Hospital, Southern Medical University, Guangzhou, China. FAU - Bai, Xiaochun AU - Bai X AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. AD - State Key Laboratory of Organ Failure Research, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China. LA - eng PT - Journal Article DEP - 20180326 PL - Germany TA - Cell Physiol Biochem JT - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology JID - 9113221 RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (Intracellular Signaling Peptides and Proteins) RN - 0 (Platelet Endothelial Cell Adhesion Molecule-1) RN - 0 (RNA, Small Interfering) RN - 0 (Vascular Endothelial Growth Factor A) RN - EC 2.7.1.1 (DEPTOR protein, human) RN - EC 2.7.10.1 (Vascular Endothelial Growth Factor Receptor-2) RN - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1) RN - W36ZG6FT64 (Sirolimus) SB - IM MH - Animals MH - Aorta/metabolism/pathology MH - Down-Regulation/drug effects MH - Human Umbilical Vein Endothelial Cells MH - Humans MH - Hypoxia-Inducible Factor 1, alpha Subunit/metabolism MH - Intracellular Signaling Peptides and Proteins/antagonists & inhibitors/genetics/*metabolism MH - Kidney/metabolism/pathology MH - Liver/metabolism/pathology MH - Mechanistic Target of Rapamycin Complex 1/antagonists & inhibitors/*metabolism MH - Mice MH - Mice, Knockout MH - *Neovascularization, Physiologic/drug effects MH - Platelet Endothelial Cell Adhesion Molecule-1/metabolism MH - RNA Interference MH - RNA, Small Interfering/metabolism MH - Signal Transduction/drug effects MH - Sirolimus/pharmacology MH - Up-Regulation/drug effects MH - Vascular Endothelial Growth Factor A/genetics/metabolism MH - Vascular Endothelial Growth Factor Receptor-2/metabolism OTO - NOTNLM OT - Angiogenesis OT - DEPTOR OT - HIF-1alpha OT - VEGF OT - Vascular endothelial cell OT - mTORC1 EDAT- 2018/04/04 06:00 MHDA- 2018/07/06 06:00 CRDT- 2018/04/04 06:00 PHST- 2017/08/24 00:00 [received] PHST- 2018/02/21 00:00 [accepted] PHST- 2018/04/04 06:00 [pubmed] PHST- 2018/07/06 06:00 [medline] PHST- 2018/04/04 06:00 [entrez] AID - 000488619 [pii] AID - 10.1159/000488619 [doi] PST - ppublish SO - Cell Physiol Biochem. 2018;46(2):520-531. doi: 10.1159/000488619. Epub 2018 Mar 26.